FlowCellect Rodent NSC Characterization Kit (Astrocyte)

FlowCellect Rodent NSC Characterization Kit (Astrocyte)

FlowCellect

25 Tests / Kit

Millipore’s The FlowCellect™Rodent Neural Stem Cell Characterization Kit for Astrocyte Differentiation is designed as an easier and quicker way to track Astrocyte differentiation. The kit will also help to determine the percentage of undifferentiated rat neural stem cells in culture by cells that express both Sox-2 and Nestin as well as track GFAP upregulation upon Astrocyte differentiation. This quick test will allow researchers to test the quality of their cells in culture as well as see changes in marker expression during a differentiation protocol.

Sufficient reagents are provided for 25 3-color samples and 5 isotype control tests. The kit includes all optimized fluorescently labeled antibodies and buffers necessary for analysis. To verify antibody specificity, all antibodies were tested against Rat Neural Stem Cells as well as Rat Astrocytes to verify antibody specificity. A cell strainer is also included to ensure samples are devoid of large cell clumps that can alter cell analysis. Detailed assay instructions are also included to assist in analysis and to ensure the correct cell concentration is obtained for flow cytometric analysis.

Rat Neural Stem cells (rat NSCs) can be maintained in vitro as self renewing multi-potent cells that can be induced to differentiate into astrocytes, neurons, or oligodendrocytes by manipulating culture conditions (1). Therefore, Rat NSCs have the potential of providing an unlimited supply of cells to study differentiation of NSCs to neuronal cell types. Due to the host of neurodegenerative diseases a vast amount of research is generated to study the promise of potential therapeutics involving NSCs.

Currently, the most utilized method to determine the undifferentiated state of Rat Neural Stem Cells is to analyze the expression of Sox2 and Nestin (3,4). Neural progenitor cells expressing both Sox2 and Nestin have been shown to self-replicate as well as give rise to astrocytes, neurons, and oligodendrocytes under the correct culture conditions (3, 4). GFAP is an astroglia-specific intermediate filament protein (5). Upon differentiation to an astrocyte lineage, Sox2 and Nestin expression levels decrease and GFAP expression increases.

The FlowCellect™ Rat Neural Stem Cell Characterization kit for Astrocytes is designed for cell biology applications. The kit uses an antibody detection method, utilizing fluorescently conjugated primary antibodies to detect Sox2, Nestin and GFAP. The FlowCellect™ Rat Neural Stem Cell Surface Expression kit is optimized for flow cytometers with a single blue (488 nm) laser light source.

The kit is optimized for use on the Guava EasyCyte™ Plus cytometer that requires cell concentrations to be less than 500 cells per microliter. The kit does not include culture media or other materials that should be readily available in a common laboratory setting that routinely performs experiments of the type described here.

• Flow Cytometry
• in vitro Toxicology Assays

Rat

• Unless otherwise stated in our catalog or other company documentation accompanying the product(s), our products are intended for research use only and are not to be used for any other purpose, which includes but is not limited to, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses or any type of consumption or application to humans or animals.
• Alexa Fluor® is a registered trademark of Molecular Probes, Inc.

Lot to lot in-process testing on all kit components

Neural Stem Cells

Stem Cell Research

Characterization

• 1. 10X Wash buffer: (Part No. CS202123) 1 bottle containing 13mL
• 2. Fixation buffer: (Part No. CS202122) 1 bottle containing 13mL
• 3. 5X Assay buffer: (Part No. CS202124) 1 bottle containing 55mL
• 4. 1X Permeabilization Buffer: (Part No. CS202125) 1 bottle containing 13mL
• 5. 20X IgG2A-FITC isotype control-(Catalog No. CS202584)
• 6. 20X IgG1-PE isotype control-(Catalog No. CS202573)
• 7. 20X IgG1-PE/CY5 isotype control-(Catalog No. CS202595)
• 8. 20X Sox-2 FITC-(Catalog No. CS202594)
• 9. 20X Nestin-PE-(Catalog No. CS202593)
• 10. 20X GFAP-PE/CY5-(Catalog No. CS202592)
• 11. Steri-flip cell strainer (60mm): (Catalog # SCNY00060)

• P48431
• P14136

4-8°C; 6 Months

Mouse

• Fluorescent
• ¹²⁵I

• SOX2
• NES
• GFAP

FlowCellect Rodent NSC Characterization Kit (Astrocyte)

• NM_003106.2
• NM_002055.2

Flow Cellect

ExpressPro

Monoclonal Antibody

Research